KR20240030971A - Method and system for processing meat using probiotic-based antimicrobial agents - Google Patents

Abstract

The present invention performs sterilization treatment on meat carcasses using kimchi lactic acid bacteria-derived antibacterial substances, thereby extending the storage period by suppressing the growth of spoilage microorganisms, as well as maintaining quality and palatability, such as changes in meat color and water holding capacity and prevention of fat rancidity. High discloses an antibacterial treatment system for meat slaughter and an antibacterial treatment method using the same.

Description

{Method and system for processing meat using probiotic-based antimicrobial agents}

The present invention relates to a meat processing method and system using lactic acid bacteria-based antibacterial substances.

Livestock products can easily deteriorate due to unsanitary conditions during the slaughter process and poor supply and demand processes during the distribution process. The production of livestock products can be infected by pathogenic microorganisms throughout the process from slaughter to consumer.

In particular, unavoidable contamination by microorganisms inherent in the working environment, workers, work tools, and work benches during the slaughter process is analyzed as a major factor in the spoilage and quality deterioration of fresh meat during the distribution period.

Livestock products are more prone to spoilage or deterioration than other foods because they contain livestock blood, moisture, and various nutrients, which leads to rapid growth of spoilage microorganisms. Therefore, an appropriate processing method is needed to distribute and sell livestock products.

Meat carcasses obtained from livestock products are immediately sealed and packaged for distribution and undergo a heat treatment process for sterilization. The remaining juices and oils that escape from this heat treatment process are distributed along with the meat slaughter, shortening the product's shelf life and deteriorating its quality. In addition, heat treatment or preservative treatment has been performed to extend the shelf life, but it is true that these existing methods not only have a high probability of destroying nutrients, but also increase consumer concerns about the safety of additives.

In addition to the above heat treatment, methods of sterilizing meat carcasses by spraying a sterilizing liquid such as alcohol or irradiating radiation are used, but these methods alone are insufficient to obtain a sufficient effect.

The present invention focuses on kimchi lactic acid bacteria and provides a method and system for processing meat using them in order to obtain a high antibacterial effect without affecting the quality of meat carcasses.

Kimchi lactic acid bacteria are lactic acid bacteria generated during the fermentation process of kimchi and are effective in eliminating harmful microorganisms. There are about 200 types of microorganisms in kimchi fermentation, but the main lactic acid bacteria include 3 genera and about 20 species, of which the commonly known genus Lactobacillus and Leuconostoc Lactic acid bacteria of the genus Leuconostoc and Weissella are the main species.

New species of lactic acid bacteria recently discovered in kimchi include Leuconostoc Kimchii , Leuconostoc inhae , and Weissella confusa, and these lactic acid bacteria have antioxidant, anti-aging, anti-cancer, and anti-oxidant properties. Its nutritional importance and various functionalities, including immunomodulatory functions, have been studied, and research on microorganisms involved in the fermentation stage is also ongoing.

Kimchi lactic acid bacteria are mainly applied to the food field, but, as in Patent Documents 1 to 3, their use in cosmetic compositions using the antibacterial effect of the kimchi lactic acid bacteria has also been proposed.

The present invention was intended to apply kimchi lactic acid bacteria to the antibacterial effect of meat carcasses. Looking at Patent Document 4, a method of processing livestock products using lactic acid bacteria is disclosed, but the types of lactic acid bacteria are Enterococcus faecalis , Lactobacillus plantarum , and Lactobacillus casei . , and Lactobacillus acidophilus , which are different types of lactic acid bacteria from kimchi lactic acid bacteria. Additionally, this patent does not directly demonstrate the antibacterial effect of applying the lactic acid bacteria.

KR public 10-2007-0089474 (2007.08.31) KR public 10-2020-0084829 (2020.07.13) KR Registration 10-2125316 (2020.06.16) KR public 10-2022-0071772 (2022.05.31)

The rich nutrients present inside the meat serve as an appropriate source of nutrients for the growth of contaminated microorganisms during the slaughter and processing process, thereby promoting the spoilage of the meat. As the storage period after slaughter passes, the number of microorganisms gradually increases, and when it exceeds a certain level, spoilage occurs.

There is a way to lower the initial microbial count in order to preserve slaughtered meat for a long time. The present invention applies kimchi lactic acid bacteria, a type of lactic acid bacteria, to the antibacterial treatment process of meat carcass to reduce the initial number of microorganisms in the slaughtered meat, suppress the subsequent growth of microorganisms, and at the same time increase the effectiveness of the meat. A composition was prepared that did not affect quality.

The purpose of the present invention is to provide an antibacterial composition for meat slaughter and antibacterial treatment using the same.

In order to achieve the object of the present invention, the present invention provides an antibacterial composition for meat slaughter containing a pro-bacterial substance derived from kimchi lactic acid bacteria as an active ingredient.

As used herein, ‘meat carcass (carcass)’ refers to slaughtered meat (carcass) of edible mammals and poultry.

The mammals include cows, pigs, sheep, horses, goats, deer, rabbits, etc., and the poultry include chickens, ducks, geese, turkeys, etc.

The antibacterial composition for meat carcasses of the present invention was able to lower the initial number of microorganisms for long-term preservation of slaughtered meat, and to inhibit microbial growth and secure antibacterial effects without deteriorating the quality of meat carcasses.

Kimchi lactic acid bacteria appear during the fermentation process of kimchi, a product made by pickling radish, cabbage, cucumber, etc. in salt, or mixing the pickled material with seasonings such as red pepper, garlic, green onion, ginger, and salted fish, then maturing by producing lactic acid and fermenting at low temperature. It is a kimchi lactic acid bacteria.

The pro-bacterial material derived from kimchi lactic acid bacteria is at least one selected from the group consisting of a culture solution of kimchi lactic acid bacteria, a concentrate of the culture solution, a dried product of the culture solution, and mixtures thereof. At this time, the culture medium of kimchi lactic acid bacteria is a filtrate obtained by filtering or centrifuging the culture medium to remove strains, or a supernatant obtained by centrifuging the culture medium, sonicating the culture medium, or cells obtained by treating the culture medium with a lysozyme. It is one or more types selected from crushed fluids.

In the antibacterial composition for meat slaughter of the present invention, the antibacterial material derived from kimchi lactic acid bacteria is preferably contained in the range of 0.001 to 99.9% by weight, 0.001 to 70% by weight, 0.01 to 60% by weight, and 0.1 to 50% by weight in the total composition. . At this time, if the content of the antibacterial substance is less than the above range, it may be difficult to obtain the expected effect, and even if it exceeds the above range, the effect may not be significantly increased by the amount added.

The kimchi lactic acid bacteria according to the present invention as described above may be any one selected from the genus Lactobacillus , kimchi lactic acid bacteria of the genus Leuconostoc , kimchi lactic acid bacteria of the genus Weissella , and mixed lactic acid bacteria thereof. there is.

Kimchi lactic acid bacteria of the Lactobacillus genus include Lactiplantibacillus plantarum , Lactilactobacillus sakei , Lacticaseibacillus paracasei, and Lactobacillus algidus . , Lactobacillus brevis , Lactobacillus curvatus , Lactobacillus kimchii, Lactobacillus mali , Lactobacillus pentosus , Lactobacillus plantarium. plantarum ), Lactobacillus sakei, Lactobacillus casei , Lactobacillus paracasei , etc.

Kimchi lactic acid bacteria belonging to the genus Leuconostoc include Leuconostoc citreum, Leuconostoc lactis , and Leuconostoc mezenteroid subsp. Dextranicum ( Leuconostoc mesenteroides subsp. dextranicum ), Leuconostoc mesenteroides subsp. Mesenteroides ( Leuconostoc mesenteroides subsp. Mesenteroides ), Leuconostoc argentinum , Leuconostoc carnosum, Leuconostoc gellidum , Leuconostoc kimchii , Leuconostoc inhae , Leuconostoc gasicomitatum , etc.

Kimchi lactic acid bacteria of the Weissella genus are Weissella koreensi , Weissella hanii , Weissella kimchii , Weissella soli , and Weissella konfusa ( Weissella confusa ), etc.

Preferably, the kimchi lactic acid bacteria of the present invention may be Lactobacillus genus Kimchi lactic acid bacteria, and more preferably 2 selected from the group consisting of Lactyplantybacillus plantarium, Lactylactobacillus sakei, and Lacticajay Bacillus paracasei. It may be more than one species, more preferably a mixed strain of Lactiplantybacillus plantarium and Lacticajay Bacillus paracasei, and even more preferably 5 to 40% by weight of Lactiplantybacillus plantarium, and Lacticajay Bacillus paracasei. It may be a mixed strain containing 60 to 95% by weight of casei.

At this time, Lactiplantybacillus Plantarium may be Lactiplantybacillus Plantarium 3104 and Lactiplantybacillus Plantarium 13903, and when used as a mixed strain, Lactiplantybacillus Plantarium 3104 is 2.5 to 30% by weight within 100% by weight of the mixed strain. %, Lactiplantibacillus plantarium 13903 can be used at 2.5 to 30% by weight.

In addition, LacticaJ Bacillus paracasei may be LacticaJ Bacillus paracasei 1011, LacticaJ Bacillus paracasei 1015, and LacticaJ Bacillus paracasei 1044, and when used as a mixed strain, within 100% by weight of the mixed strain. It can be used at 10 to 40% by weight of LacticaJ Bacillus paracasei 1011, 10 to 50% by weight of LacticaJ Bacillus paracasei 1015, and 10 to 40% by weight of LacticaJ Bacillus paracasei 1044.

According to one embodiment, the kimchi lactic acid bacteria of the present invention include Lactica J Bacillus paracasei 1015 used in a high content of 50% by weight, and Lactiplantybacillus Plantarium 3104 and Lactiplantybacillus Plantarium 13903 each containing 50% by weight. Excellent results were obtained when mixed at a weight percent content.

The remainder of the antibacterial composition for meat slaughter of the present invention is water. The water may be, for example, pure water, ultrapure water, distilled water, purified water, water for injection, tap water, etc.

According to one embodiment, the antibacterial composition for meat slaughter of the present invention further includes at least one selected from the group consisting of acetic acid and ethanol.

Acetic acid itself has an antibacterial effect, but when sprayed or immersed in meat carcasses, the unique scent of acetic acid not only makes it inappropriate to use, but also reduces the quality of the meat. Ethanol has no antibacterial effect on its own.

However, in the present invention, when acetic acid and/or ethanol are used together with kimchi lactic acid bacteria, it has a superior antibacterial effect compared to when kimchi lactic acid bacteria are used alone. In particular, problems caused by acetic acid can be prevented by using acetic acid at low concentrations.

Preferably, acetic acid is used in an amount of 3% by weight or less, 2% by weight or less, and 1% by weight or less based on the total amount of pro-bacterial substances derived from kimchi lactic acid bacteria. Additionally, ethanol is used in amounts of 10% by weight or less, 7% by weight or less, and 5% by weight or less. If necessary, a mixture of the two is also possible.

The kimchi lactic acid bacteria culture medium according to the present invention is a culture medium obtained by inoculating kimchi lactic acid bacteria into a medium and culturing it at 20 to 40° C. for 1 to 4 days, or a filtrate obtained after centrifuging the culture medium.

At this time, the medium is not particularly limited and any known medium can be used, for example, one of natural medium, synthetic medium, selective medium, and differential medium. can be used These media can be prepared and used directly, or you can purchase and use commercially available media such as Nutrient Agar media and MRS agar media.

According to one embodiment, the medium of the present invention is a natural medium, and among them, it may be an edible medium (liquid).

The edible medium is prepared by using cabbage juice and cabbage juice together. When the cabbage juice and cabbage juice are mixed at a weight ratio of 1:9 to 9:1 and added to the medium, the number of cultured bacteria could be effectively increased.

The specific composition of the edible medium includes 5 to 30% by weight of cabbage juice and cabbage juice, plus 1 to 5% by weight of glucose, 0.5 to 5% by weight of corn steep liquid, 0.1 to 1% by weight of sodium acetate, and sulfuric acid. It may contain 0.001 to 0.05% by weight of manganese, with the balance being water.

Additionally, the growth effect of kimchi lactic acid bacteria can be obtained when peptone and polysorbate 80 (Tween80) are added to the edible medium. The peptone is used in an amount of 1 part by weight or less, and polysorbate 80 is used in an amount of 0.5 parts by weight or less per 100 parts by weight of cabbage and cabbage juice. According to one embodiment, when polysorbate 80 was added at 0.1 parts by weight, the most advantageous results were obtained in securing the convenience and effectiveness of medium production.

The method of antibacterial treatment of meat carcasses using the composition of the present invention is not particularly limited in the present invention, and various methods may be used.

According to one embodiment, meat carcasses are treated antibacterially by spraying an antibacterial composition for meat carcasses onto the carcasses. At this time, the antibacterial composition is diluted in water and used, and antibacterial treatment can be performed by spraying at 1 to 2 atmospheres of pressure for 5 to 60 seconds.

According to another embodiment, meat carcass is immersed in an antibacterial composition. The antibacterial composition is also used by diluting it in water. The immersion time is selectively determined depending on the type and size of the meat carcass, so it is not particularly limited, but is preferably immersed for 5 to 420 seconds.

The dilution ratio at the time of dilution may be any dilution ratio that can achieve a concentration that exerts an antibacterial effect as a composition, and is not particularly limited, but is preferably 1 to 10,000 times, more preferably 1 to 1,000 times.

Through this antibacterial treatment, the initial number of microorganisms in slaughtered meat is lowered and subsequent growth of microorganisms is suppressed, while at the same time, it does not affect the quality of the meat. The microorganisms may include E. coli , Staphylococcus aureus, Shigella flexneri , and Salmonella typhimurium , which cause meat spoilage. .

The antibacterial treatment of the present invention allows long-term storage (low temperature) for more than 25 days, or preferably more than 45 days, thereby increasing the shelf life of meat carcasses.

If bacteria or mold invade the surface or tissue of meat, it not only causes abnormalities in the appearance, odor, and color of the meat, but also causes hygienic hazards such as spoilage, making it unsuitable for human consumption. Meat carcasses have a lot of moisture, nitrogen compounds in various stages of decomposition exist together with salts and growth nutrients, and furthermore, because they contain fermentable carbohydrates and have a high pH, bacterial growth can occur quickly.

By performing antibacterial treatment on meat carcasses using the antibacterial material derived from kimchi lactic acid bacteria of the present invention, the storage period is extended by suppressing the growth of spoilage microorganisms, as well as maintenance of quality and palatability, such as changes in meat color and water holding capacity and prevention of fat rancidity. A heightening effect can be expected.

Figure 1 shows the strains to be used in the test.
Figure 2 is a photograph showing the kimchi lactic acid bacteria culture medium, which is an antibacterial material to be used in the test.
Figure 3 is a photograph showing the antibacterial effect of kimchi lactic acid bacteria and control bacteria.
Figure 4 is a graph showing the sizes of growth inhibitory rings of kimchi lactic acid bacteria and control bacteria.
Figure 5 is a photograph showing the culture of kimchi lactic acid bacteria in an edible medium.
Figure 6 is a photograph showing the culture of kimchi lactic acid bacteria for measuring the number of viable bacteria in an edible medium, and Figure 7 is a photograph showing the measurement of the number of viable cells.
Figure 8 is a photograph showing the culture of kimchi lactic acid bacteria for measuring the number of viable bacteria in an edible medium containing peptone and polysorbate 80.
Figure 9 is a photograph showing the process of performing the antibacterial activity of kimchi lactic acid bacteria according to the type of culture medium.
Figure 10 is a photograph showing bacterial count.
Figure 11 is a photograph showing the process of performing the antibacterial activity of kimchi lactic acid bacteria according to the type of culture medium.
Figure 12 is a photograph showing a sterilized kimchi lactic acid bacteria culture medium.
Figure 13 is a photograph showing a sprayer containing kimchi lactic acid bacteria culture medium.
Figure 14 shows sampling on day 0 of the strip, Figure 15 shows sampling on the 7th day, Figure 16 shows sampling on the 15th day, Figure 17 shows sampling on the 30th day, and Figure 18 shows sampling on the 45th day of the strip. Shows sampling.
Figure 19 is a photograph showing a day 0 strip tip treated with a homogenization test solution, and Figure 20 is a photograph showing a day 0 tenderloin treated with a homogenization test solution.
Figure 21 is a photograph showing the surface of a tenderloin on day 0 treated with a surface bacteria test solution.
Figure 22
Figure 23

Hereinafter, to aid understanding of the present invention, it will be described in detail through examples. However, the embodiments according to the present invention may be modified into various other forms, and the scope of the present invention should not be construed as being limited to the following embodiments. Embodiments of the present invention are provided to more completely explain the present invention to those skilled in the art.

The antibacterial material mentioned in the examples below refers to kimchi lactic acid bacteria culture medium.

Test Example 1: Selection of Kimchi Lactobacillus

(1) Strain and culture used

As shown in Figure 1, a total of 20 types (16 types of Kimchi lactic acid bacteria ( Lactiplantibacillus plantarum, Latilactobacillus sakei, Lacticaseibacillus paracasei )) and 4 types of pathogens used in the present invention were prepared.

After preparing 14 types of kimchi lactic acid bacteria cultures cultured in 15ml of MRS Broth (Difco, USA), 1.5ml of each strain was divided into sterilized 2ml centrifuge tubes and centrifuged at 6000xg for 15 minutes to recover the culture supernatant. The obtained culture supernatant was dispensed into bottles for each type, and the remaining bacterial cells were removed from the recovered culture supernatant using a syringe filter (PES, 0.45㎛) to recover the filtrate. The recovered filtrate was adjusted to pH 7.0 and refrigerated at 4°C. Figure 2 is a photograph showing the antibacterial material (kimchi lactic acid bacteria culture medium) to be used in the test.

(2) Measurement of antibacterial effect

Four types of pathogens were prepared, cultured in 30ml of Nutrient Broth (NB; Difco, USA) at 37°C for 48 hours, divided into sterilized 2ml centrifuge tubes, divided into 2ml of each strain, centrifuged at 6000×g for 15 min, and the culture supernatant was removed. . Next, the precipitated strains were suspended in 2 ml of sterilized distilled water and homogenized by inoculating each strain into 300 ml of Nutrient agar stored in a constant temperature water bath at 45°C. Then, in a clean bench maintained aseptically, 11 plates for each strain were made in a Petri dish (total of 44P), hardened, placed on an 8mm paper disc (Advantec, Japan), and 50 μl of the prepared kimchi lactic acid bacteria and control group were dispensed each. Then, it was sealed with parafilm and cultured in an incubator at 37°C for 18 hours. After culture, when antibacterial activity was confirmed as shown in Figure 3, the size of the ring formed was measured using a vernier caliper.

(3) Antibacterial activity results

The antibacterial effect of kimchi lactic acid bacteria is shown in the table below.

The above results show that acetic acid and kimchi lactic acid bacteria have antibacterial activity.

(4) Antibacterial activity statistics

Referring to Table 1 above, antibacterial tests were performed three times under the same conditions, and the average size of growth inhibitory rings is shown in Table 2 and Figure 4 below. Figure 4 is a graph showing the sizes of growth inhibitory rings of kimchi lactic acid bacteria-derived pro-bacterial substances and control bacteria.

Looking at the table above, among kimchi lactic acid bacteria, Lb. The average size of growth inhibitory rings of paracasi kimchi lactic acid bacteria was the highest, and its shape was also confirmed to be distinct. The size of the rings produced by the Lb.plantarum species was large, but there were few strains that showed a distinct shape like the Lb.paracasei species.

In the case of ethanol, even if the concentration was increased, there was no effect.

Acetic acid had the best growth inhibition ability, but is expected to cause an unpleasant odor when applied to meat, and is expected to affect protein and fat due to the acid component.

(5) Selection of kimchi lactic acid bacteria

Based on the above results, kimchi lactic acid bacteria with excellent antibacterial activity were finally selected.

Test Example 2: Antibacterial activity of edible medium and kimchi lactic acid bacteria

This experiment was performed to check the activity of kimchi lactic acid bacteria when making a medium with edible materials.

(1) Manufacturing edible medium

A medium using cabbage juice and cabbage juice was prepared with the composition shown in the table below. At this time, purified water was used for the remaining composition, where % is weight %. The edible medium prepared after mixing each composition was sterilized at 120°C and 2 atm for 15 minutes, and then for the experiment, 30 ml of the edible medium was dispensed into sterilized 50 ml conical tubes. In the table below, cabbage refers to cabbage juice, and cabbage refers to cabbage juice.

(2) Kimchi lactic acid bacteria inoculation and liquid culture

Kimchi lactic acid bacteria stored frozen were thawed and used. After thawing, centrifugation was performed, the supernatant was discarded, 1.5 ml of sterilized water was added, vortexed, and 500 μl each was inoculated into a conical tube according to the addition ratio of cabbage and cabbage. After vortexing using a vortex mixer, the conical tube lid was sealed using parafilm, and cultured in an incubator at 30°C for 48 hours. Figure 5 is a photograph showing the culture of kimchi lactic acid bacteria in an edible medium.

(3) Measurement of viable cell count

1L of MRS agar (Difco, USA) was prepared and sterilized, and the temperature of the water bath was maintained at 50°C. Kimchi lactic acid bacteria that had been liquid-cultured in an edible medium were vortexed using a vortex mixer, and then 1 ml each was extracted from the culture solution to prepare 10 ¹ and diluted to 10 ⁹ . Then, the 10 ⁶ to 10 ⁹ diluted solution was repeatedly dispensed twice, 1 ml each, into a 90 mm Petri dish. For this purpose, 20 ml of the prepared MRS agar (Difco, USA) medium was dispensed. After mixing by shaking carefully to avoid touching the lid and sides of the Petri dish, it was sealed using parafilm and cultured in an incubator at 30°C for 48 hours.

(4) Results

Figure 6 is a photograph showing the culture of kimchi lactic acid bacteria for measuring the number of viable bacteria in an edible medium, and Figure 7 is a photograph showing the measurement of the number of viable cells. After going through this process, the table below shows the number of viable bacteria measured according to the content of cabbage and cabbage juice.

Looking at the above results, the highest viable bacterial count was measured on average when cabbage and cabbage were mixed at a ratio of 1:1, showing that the mixed use is more effective in proliferating the viable bacterial count than single items. Based on these results, the most desirable edible medium composition is shown in the table below.

Test Example 3: Selection of edible medium additives

This was conducted to check whether peptone and polysorbate 80, which are known to be useful for the growth of lactic acid bacteria, were added to the edible medium.

(1) Production of edible medium

Edible medium was prepared with the composition shown in the table below. In this case, % is weight%.

(2) Kimchi lactic acid bacteria inoculation

*Frozen stored kimchi lactic acid bacteria were thawed and used. After thawing, centrifugation was performed, the supernatant was discarded, 2.0 ml of sterilized water was added, vortexed, and 500 μl each was inoculated into a 250 mL Erlenmeyer flask according to the addition ratio of cabbage and cabbage. After vortexing using a vortex mixer, the mouth of the flask was sealed using parafilm, and cultured in an incubator at 30°C for 48 hours. Figure 5 is a photograph showing the culture of kimchi lactic acid bacteria in an edible medium.

(3) Results

Figure 8 is a photograph showing the culture of kimchi lactic acid bacteria for measuring the number of viable bacteria in an edible medium to which peptone and polysorbate 80 were added, and the table below shows the measured number of viable cells.

Looking at the table above, it can be seen that adding peptone and polysorbate 80 to the edible medium is effective for the growth of kimchi lactic acid bacteria. However, in the case of peptone, addition of more than 1% did not show a significant effect on growth, and in the case of polysorbate 80, addition of 0.1% was found to be most effective.

Test Example 4: Antibacterial activity of kimchi lactic acid bacteria according to medium type

An antibacterial treatment process was added to the antibacterial material that had been sterilized and compared with an edible medium without antibacterial treatment and MRS medium to determine the antibacterial effect.

(1) Materials and experimental methods

- Kimchi lactic acid bacteria culture medium made from edible medium

250 ml of the kimchi lactic acid bacteria culture solution, which is each antibacterial material, was dispensed into sterilized 500 ml glass bottles. The autoclave was set to 65℃, the center temperature was heated to 65℃, and then sterilized for 30 minutes. After sterilization, the sample was aseptically transferred to a 15ml conical tube on a clean bench.

(2) Antibacterial experiment

250 ml of Nutrient agar (NA; Difco, USA) medium was prepared in a 500 ml Erlenmeyer flask and sterilized at 121°C and 2 atm for 15 minutes. The sterilized 500ml Erlenmeyer flask was cooled in a water bath at 45°C. Afterwards, the pathogens that had been frozen were taken out, added to the cooled NA medium, and stirred for 5 minutes on a stirrer. After stirring was completed, the NA mixed with each pathogen was solidified by dispensing 20 ml into 90 mm Petri dishes. A sterilized 8 mm paper disc (Advantec. Japan) was placed on the hardened NA medium, and 50 μl of each strain was dispensed and cultured in an incubator at 37°C for 24 hours.

Detection and confirmation of the number of general bacteria in the kimchi lactic acid bacteria culture medium, which is three types of antibacterial substances, was performed. After sampling of the kimchi lactic acid bacteria culture medium, which is each antibacterial material, was dispensed at 1 ml volume onto 3M Petri film (Aerob count; AC, USA) without dilution. Afterwards, it was cultured in an incubator at 37°C for 24 hours.

(3) Results

Figure 9 is a photograph showing the process of performing the antibacterial activity of the antibacterial material according to the type of culture medium, and Figure 10 is a photograph showing the bacterial count. The table below shows the results.

Looking at the table above, MRS medium was the best in terms of the size of the inhibitory ring produced by each kimchi lactic acid bacteria, but sterilized and non-sterilized edible media also showed excellent results. This shows that it exhibits effective antibacterial activity even without performing a sterilization process when producing edible media.

Test Example 5: Antibacterial activity according to mixing ratio of kimchi lactic acid bacteria

In order to maximize the effect of suppressing harmful bacteria that may affect meat, this experiment was performed to find the optimal ratio by mixing kimchi lactic acid bacteria culture medium, which is a pre-bacterial substance called kimchi lactic acid bacteria.

(1) Materials

Through the above antibacterial experiment, the most effective strain was selected, and its contents were mixed as follows to prepare a mixed strain. In the table below, % is weight percent.

(2) Antibacterial test

250 ml of Nutrient agar (NA; Difco, USA) medium was prepared in a 500 ml Erlenmeyer flask and sterilized at 121°C and 2 atm for 15 minutes. The sterilized 500ml Erlenmeyer flask was cooled in a water bath at 45°C. Afterwards, the pathogens that had been frozen were taken out, added to the cooled NA medium, and stirred for 5 minutes on a stirrer. After stirring was completed, the NA mixed with each pathogen was solidified by dispensing 20 ml into 90 mm Petri dishes. A sterilized 8 mm paper disc (Advantec. Japan) was placed on the hardened NA medium, and 50 μl of each strain was dispensed and cultured in an incubator at 37°C for 24 hours.

(3) Results

Figure 11 is a photograph showing the process of performing the antibacterial activity of kimchi lactic acid bacteria according to the type of culture medium, and the table below shows the results.

Looking at the table above, it can be seen that all of the mixed strains 1 to 5 have antibacterial activity. Among them, a mixture of Lb.paracasei 1011 and Lb.paracasei 1015, and Lb.plantarum 3104 and Lb.plantarum 13093 showed excellent results.

Test Example 6: Antibacterial activity according to the type and mixing ratio of additional ingredients

In Table 1, acetic acid, which has antibacterial activity, and ethanol, which does not have antibacterial activity, were used to provide storage properties for antibacterial substances and conduct this experiment to determine antibacterial efficacy.

(1) Materials

Through the above antibacterial experiment, the most effective strain was selected, and an antibacterial composition was prepared by adding acetic acid and ethanol, respectively. In the table below, the % of kimchi lactic acid bacteria is weight %, and acetic acid and ethanol are weight % based on the total amount of kimchi lactic acid bacteria.

(2) Antibacterial experiment

250 ml of Nutrient agar (NA; Difco, USA) medium was prepared in a 500 ml Erlenmeyer flask and sterilized at 121°C and 2 atm for 15 minutes. The sterilized 500ml Erlenmeyer flask was cooled in a water bath at 45°C. Afterwards, the pathogens that had been frozen were taken out, added to the cooled NA medium, and stirred for 5 minutes on a stirrer. After stirring was completed, the NA mixed with each pathogen was solidified by dispensing 20 ml into 90 mm Petri dishes. A sterilized 8 mm paper disc (Advantec. Japan) was placed on the hardened NA medium, and 50 μl of each strain was dispensed and cultured in an incubator at 37°C for 24 hours.

(3) Results

The table below shows the antibacterial activity of the antibacterial composition according to the addition of acetic acid and ethanol.

Looking at the table above, all tests in which acetic acid was added and tests 4 and 5 in which ethanol was added showed excellent antibacterial activity, and it can be seen that the addition of these substances helps the antibacterial activity.

Test Example 7: Antibacterial effect according to antibacterial treatment on meat

This experiment was performed to confirm the effect of inhibiting microbial growth when processing meat with the antibacterial material containing the kimchi lactic acid bacteria culture medium of the present invention.

(1) Mass cultivation of kimchi lactic acid bacteria

Kimchi lactic acid bacteria culture medium, which had been cultured for 48 hours in an incubator at 30°C, was prepared and centrifuged at 2500xg for 30 minutes to separate the bacterial cells and the culture supernatant. The obtained culture supernatant was dispensed into bottles for each type, and the remaining bacterial cells were removed from the recovered culture supernatant using a syringe filter (PES, 0.45㎛) to recover the filtrate. The recovered filtrate (antibacterial material) was adjusted to pH 7.0 and refrigerated at 4°C.

Figure 12 is a photograph showing a sterilized kimchi lactic acid bacteria culture medium.

(2) Material preparation

We prepared two parts of meat (tenderloin and strip tip), basically removed unnecessary fat and fascia, and prepared approximately 400-450g each. Each antibacterial material was filled into a spray bottle that had been washed in boiling water, sprayed to the extent that the surface of the meat was wet, then vacuum-packed and stored at 2°C. At this time, Figure 13 is a photograph showing a sprayer in which a culture solution is stored for use in processing meat.

Figure 14 shows sampling on day 0 of the strip, Figure 15 shows sampling on the 7th day, Figure 16 shows sampling on the 15th day, Figure 17 shows sampling on the 30th day, and Figure 18 shows sampling on the 45th day of the strip. Shows sampling. Sampling of tenderloin and tenderloin (surface) was conducted in the same manner. At this time, the abbreviations are as shown in the table below.

(3) Preparation of test solution

To measure the antibacterial effect, a homogenization test solution was prepared for the strip tip and tenderloin, and a surface bacteria test solution was prepared for the surface of the tenderloin using a cotton swab.

- Preparation of homogenization test solution -

Meat samples sprayed with antibacterial substances corresponding to day 0 and control meat samples were prepared. The prepared specimens were aseptically separated into sterile bags of 25 g each using sterilized scissors and forceps. A test solution was prepared by adding 225 ml of sterilized saline solution (0.85%, NaCl) and sufficiently homogenizing it using a tool. 1ml of the prepared test solution was added to 9ml sterilized saline solution as a stock solution, vortexed, and diluted 10 times to prepare the solution up to 10 ^-4 .

*Figure 19 is a photograph showing that day 0 strip tip was treated with a homogenization test solution, and Figure 20 is a photograph showing a day 0 tenderloin treated with a homogenization test solution.

- Preparation of surface bacteria test solution -

Meat samples sprayed with antibacterial substances corresponding to day 0 and control meat samples were prepared. The surface of the sample (10cm The prepared 10-fold diluted test solution was prepared up to 10 ^-4 .

Figure 21 is a photograph showing the surface of a tenderloin on day 0 treated with a surface bacteria test solution.

(4) Measurement of bacterial count

The general bacterial count and E. coli count were measured respectively.

-Measurement of general bacterial count-

Each sample was labeled on 3M Petri film (dry film, AC). Inside the clean bench, the cover of the Petri film was lifted upward, and 1 ml of the test solution prepared above was dispensed into the center of the medium. The cover was then gently closed and gently pressed with a pressure plate to inoculate the test solution. The test solution was repeatedly inoculated twice from 10 ^-1 to 10 ^-4 . The inoculated Petri films were overlapped for each sample and cultured in an incubator at 37°C for 48 hours, and then calculated as the average number of bacterial cells on two Petri films per sample.

-Measurement of E. coli count-

Each sample was labeled on 3M Petri film (dry film, EC). Inside the clean bench, the cover of the Petri film was lifted upward, and 1 ml of the test solution prepared above was dispensed into the center of the medium. The cover was then gently closed and gently pressed with a pressure plate to inoculate the test solution. The test solution was repeatedly inoculated twice from 10 ^-1 to 10 ^-2 . The inoculated Petri films were overlapped for each sample and cultured in an incubator at 37°C for 48 hours, and then calculated as the average number of bacterial cells on two Petri films per sample.

(5) Results

The table below shows the number of common bacteria (AC) and Escherichia coli ( E.Coli ) present on the surface of the strip, tenderloin, and tenderloin on day 0.

The shelf life of meat is influenced by the initial microorganisms, which appear to be about 10 ² to 10 ³ after slaughter, and their number increases as the storage period increases. When the number of microorganisms in meat exceeds 10 ⁶ , spoilage begins, and when it reaches 8 to 9 log CFU/㎤, a putrefactive odor occurs. In particular, harmful microorganisms proliferate from the surface and spread throughout the meat, causing spoilage.

Looking at the table above, it can be seen that when antibacterial treatment is performed with kimchi lactic acid bacteria, E. coli does not exist on the surface of meat after slaughter, and therefore, additional antibacterial treatment process can be replaced.

Figure 22 shows the carcass antibacterial liquid treatment system according to the present invention.

As an embodiment of the present invention, a system for coating meat carcasses by spraying an antibacterial liquid will be described.

As shown in Figure 22, carcass processed in unit quantities for meat products is transported to the carcass reception room of Figure 22 through the carcass reception room. The carcass is stored in the carcass storage unit 20 of the carcass storage room. The carcass storage unit can be stored by hanging the carcass so that it is exposed to the air, or placed on a perforated shelf so that the sprayed antibacterial solution can reach it.

The antibacterial composition produced in the manner described above is stored in the antibacterial liquid storage unit 11 and connected to the high pressure pump 12 and the antibacterial liquid spray nozzle 14 through a pipe equipped with a nozzle. The high pressure pump 12 generates and supplies high pressure to transfer the antibacterial composition from the antibacterial liquid storage unit 11 to the antibacterial liquid spray nozzle 14, and the antibacterial liquid spray nozzle 14 is controlled by the control unit 13. By opening and closing the nozzle, the antibacterial composition is sprayed onto the carcass.

Using this antibacterial liquid treatment system, the antibacterial composition described above is sprayed on and coated on carcass through an antibacterial liquid generation process of generating an antibacterial liquid in carcass, and a carcass spraying process in which the generated antibacterial liquid is sprayed onto the carcass for coating. The processed carcass is treated with antibacterial agents.

Claims (9)

A system for coating carcass by spraying an antibacterial solution,
A carcass storage department that stores carcass;
An antibacterial liquid storage unit that stores the antibacterial liquid;
A high pressure pump that generates high pressure to transfer the antibacterial liquid to the antibacterial liquid spray nozzle;
Antibacterial liquid spray nozzles disposed at predetermined intervals on the carcass of the carcass storage unit;
A carcass antibacterial liquid processing system comprising: According to paragraph 1,
The antibacterial liquid contains antibacterial substances derived from kimchi lactic acid bacteria as an active ingredient,
The kimchi lactic acid bacteria include 5 to 40% by weight of Lactiplantibacillus plantarum and Lactobacillus paracasei . An antibacterial composition for meat slaughter, which is a mixed strain mixed at 60 to 95% by weight;
A carcass antibacterial liquid processing system characterized by According to paragraph 2,
The antibacterial agent is one or more selected from the group consisting of kimchi lactic acid bacteria culture, concentrate of the culture, dried product of the culture, and mixtures thereof. According to paragraph 3,
The culture medium of the kimchi lactic acid bacteria is a filtrate obtained by filtering or centrifuging the culture medium to remove strains, or a supernatant obtained by centrifuging the culture medium, sonicating the culture medium, or cells obtained by treating the culture medium with a lysozyme. A carcass antibacterial liquid treatment system characterized by one or more types selected from the shredded liquid. According to paragraph 1,
The antibacterial solution is,
Contains antibacterial substances derived from kimchi lactic acid bacteria as an active ingredient,
It further includes at least one selected from the group consisting of acetic acid, ethanol, and combinations thereof,
The kimchi lactic acid bacteria are a mixed strain of Lactiplantibacillus plantarum and Lactobacillus paracasei, an antibacterial composition for meat slaughter. In the carcass antibacterial treatment method of coating carcass with an antibacterial solution,
An antibacterial liquid production process that generates a carcass antibacterial liquid;
A carcass spraying process of coating the carcass by spraying the generated antibacterial liquid on the carcass;
Includes,
The antibacterial liquid contains antibacterial substances derived from kimchi lactic acid bacteria as an active ingredient,
The kimchi lactic acid bacteria include 5 to 40% by weight of Lactiplantibacillus plantarum and Lactobacillus paracasei . An antibacterial composition for meat slaughter, which is a mixed strain mixed at 60 to 95% by weight;
An antibacterial treatment method for carcass characterized by: According to clause 6,
The antibacterial material is one or more selected from the group consisting of kimchi lactic acid bacteria culture, concentrate of the culture, dried product of the culture, and mixtures thereof. In clause 7,
The culture medium of the kimchi lactic acid bacteria is a filtrate obtained by filtering or centrifuging the culture medium to remove strains, or a supernatant obtained by centrifuging the culture medium, sonicating the culture medium, or cells obtained by treating the culture medium with a lysozyme. An antibacterial treatment method for carcass characterized by using at least one selected from among the shredded liquids. According to clause 6,
The antibacterial solution is,
Contains antibacterial substances derived from kimchi lactic acid bacteria as an active ingredient,
It further includes at least one selected from the group consisting of acetic acid, ethanol, and combinations thereof,
The kimchi lactic acid bacteria are a mixed strain of Lactiplantibacillus plantarum and Lactobacillus paracasei;
An antibacterial treatment method for carcass characterized by: